U.S. patent application number 12/597867 was filed with the patent office on 2010-06-03 for use of an active principle originating from amaranth (amaranthus) for preparing a composition intended to activate cellular energy and to protect the skin from oxidative damage.
This patent application is currently assigned to Societe D'Extraction Des Principes Actifs S.A. (ISP Vincience). Invention is credited to Jean-Marie Botto, Claude Dal Farra, Nouha Domloge.
Application Number | 20100137186 12/597867 |
Document ID | / |
Family ID | 38872497 |
Filed Date | 2010-06-03 |
United States Patent
Application |
20100137186 |
Kind Code |
A1 |
Dal Farra; Claude ; et
al. |
June 3, 2010 |
USE OF AN ACTIVE PRINCIPLE ORIGINATING FROM AMARANTH (AMARANTHUS)
FOR PREPARING A COMPOSITION INTENDED TO ACTIVATE CELLULAR ENERGY
AND TO PROTECT THE SKIN FROM OXIDATIVE DAMAGE
Abstract
Methods of administering in a cosmetic composition or a
pharmaceutical composition, an effective amount of a peptide active
principle originating from amaranth of the species Amaranthus
hypochondriacus. The active principle is intended to activate
cellular energy and to protect the skin from oxidativ damage, and
the active principle may be used alone or in association with at
least one other active principles. The active principle may also be
used in a cosmetic treatment for protecting the skin and the
appendages from external aggressions and to combat cutaneous
aging.
Inventors: |
Dal Farra; Claude; (Opio,
FR) ; Domloge; Nouha; (Valbonne, FR) ; Botto;
Jean-Marie; (Valbonne, FR) |
Correspondence
Address: |
YOUNG & THOMPSON
209 Madison Street, Suite 500
Alexandria
VA
22314
US
|
Assignee: |
Societe D'Extraction Des Principes
Actifs S.A. (ISP Vincience)
|
Family ID: |
38872497 |
Appl. No.: |
12/597867 |
Filed: |
April 23, 2008 |
PCT Filed: |
April 23, 2008 |
PCT NO: |
PCT/FR2008/000576 |
371 Date: |
October 27, 2009 |
Current U.S.
Class: |
514/20.1 ;
530/300 |
Current CPC
Class: |
A61K 8/9789 20170801;
A61Q 19/08 20130101; A61P 17/00 20180101; A61K 36/482 20130101;
A61P 17/16 20180101; A61K 36/68 20130101; A61Q 17/04 20130101; A61K
38/011 20130101; A61K 36/482 20130101; A61K 2300/00 20130101; A61K
36/68 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
514/2 ;
530/300 |
International
Class: |
A61K 8/64 20060101
A61K008/64; C07K 2/00 20060101 C07K002/00; A61Q 19/00 20060101
A61Q019/00; A61Q 19/08 20060101 A61Q019/08 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 27, 2007 |
FR |
0703062 |
Claims
1-14. (canceled)
15. A method of increasing cellular energy and protecting skin from
oxidative damage, comprising adminstrating an effective amount of a
peptide active principle, wherein, said active principle is in a
composition, and said active principle originates from the
hydrolysis of proteins from amaranth of the species Amaranthus
hypochondriachus.
16. The method according to claim 15, wherein said active principle
contains between 0.1 and 5 g/l of compounds of a peptide
nature.
17. The method, according to claim 16, wherein said active
principle contains between 0.5 and 2 g/l of compounds of a peptide
nature.
18. The method according to claim 15, wherein said active principle
is used in a quantity representing 0.0001 and 20% of the total
weight of the composition.
19. The method according to claim 18, wherein said active principle
is used in a quantity representing 0.05 and 5% of the total weight
of the composition.
20. The method according to claim 15, wherein said active principle
is solubilized in advance in a solvent selected from the group
consisting of water, glycerol, ethanol, propylene glycol, butylene
glycol, dipropylene glycol, ethoxylated or propoxylated diglycols,
cyclic polyols, vaseline, a vegetable oil, and mixtures
thereof.
21. The method according to claim 15, said composition is in a form
adapted to topical application.
22. The method according to claim 15, wherein said composition
further comprises at least one other active agent promoting the
action of said active principle.
23. The method according to claim 22, wherein said other active
principle is selected from the group consisting of principles
having an antioxidant action, active principles stimulating the
synthesis of dermal macromolecules, and active principles
stimulating energy metabolism.
24. A compound consisting of an active peptide principle, wherein
said active principle originates from the hydrolysis of proteins
from amaranth of the species Amaranthus hypochondriacus.
25. A method of protecting skin and appendages against external
aggression comprising administering to a subject in need thereof an
effective amount of the compound accordion to claim 24 in a
cosmetic composition to protect the skin and the appendages against
external aggression.
26. A method of increasing at least one of the synthesis of
intracellular ATP, the activity or synthesis of the enzyme
transphenyl transferase, and coenzyme Q10 skin cells comprising
administering to a subject in need thereof an effective amount of
compound according to claim 24 in a cosmetic composition.
27. A method of prevention or treating skin damage and appendage
damage caused by UV radiation, comprising administering to a
subject in need thereof an effective amount of compound according
to claim 24 in a cosmetic composition.
28. A method of protecting skin and appendages against oxidative
damage comprising administering to a subject in need thereof an
effective amount of compound according to claim 24 in a cosmetic
composition.
29. A method of preventing or treating the cutaneous signs of aging
and/or photo-aging, comprising administering to to a subject in
need thereof an effective effective amount of compound according to
claim 24 in a cosmetic composition.
30. A method of preventing or combating pathologies associated with
processes of oxidation, comprising administering to a subject in
need thereof an effective effective amount of compound according to
claim 24 in a pharmaceutical composition.
31. A method of cosmetic treatment comprising topically
administering to a subject in need thereof an effective amount of
compound according to claim 24 onto the skin or the appendages to
be treated.
Description
[0001] The present invention is in the cosmetic and pharmaceutical
domain, and more particularly in the domain of dermatology. The
present invention concerns the use in a cosmetic composition or for
preparing a pharmaceutical composition, of an effective quantity of
a peptide active principle originating from amaranth of the species
Amaranthus hypochondriacus, said active principle, or a composition
containing it, being intended to activate cellular energy and to
protect the skin from oxidative damage. The active principle may be
used alone or in association with at least one other active
principle. The invention also concerns a cosmetic-treatment
procedure intended to protect the skin and the appendages from
external aggressions and to combat cutaneous aging. The said active
principle can also be used to prepare pharmaceutical compositions
intended to prevent or combat pathologies linked to oxidation
processes or even certain pathologies of aging.
[0002] The term "appendages" according to the invention encompasses
the assemblage of keratinic appendices exhibited on the body
surface, in particular the hair, eyelashes, eyebrows, nails, and
hair.
[0003] The skin is a vital organ that covers the entire surface of
the body and provides protective, sensitive, immune, metabolic, or
even thermoregulatory functions. The skin, like other organs, is
subject to aging. So, one of the major mechanisms implicated in the
processes of aging is the accumulation of oxidative damage in
essential molecules such as membrane lipids, proteins, DNA, and
most particularly mitochondrial DNA (DNAmt).
[0004] Oxidative damage is caused by free radicals, chemically
unstable and very reactive species generated by intracellular
metabolism or external aggressions. Among these external
aggressions, UV rays, toxins, atmospheric pollutants, and
alimentary oxidants may be cited. Premature aging is observed in
the skin, occurring in areas exposed to radiation, characterized by
phenomena of alterations in the macromolecules (lipid peroxidation,
carbonylation of proteins) affecting, in particular, elastin,
collagen, and fibronectin. Progressive decline with age can also be
shown in the mitochondrial functions, probably linked to the
accumulation of mutations on DNAmt (K. Singh (2004), Ann. NY Acad.
Sci., 1019).
[0005] One of the important consequences of the accumulation of
oxidative damage is the reduction in the capacity of the cell to
produce ATP (Porteous et al. (1998), Eur. J. Biochem. 257(1),
192-201). Thus, the phenomenon of cellular aging is in proportion
to the oxidative damage the cell undergoes, as well as to the
process of producing the energy the cell needs to survive.
[0006] The body possesses defense mechanisms capable of trapping or
of transforming free radicals (enzymes, glutathione, vitamins A and
E, coenzyme Q10, etc.). However, these antioxidant defense systems
often prove to be insufficient under the numerous stresses and
external aggressions to which the body, and the skin in particular,
are subjected.
[0007] In this context, the particular properties of coenzyme Q10
appear to be particularly interesting.
[0008] Coenzyme Q10 (or ubiquinone) is a coenzyme present in the
mitochondrial complexes implicated in oxidative phosphorylation
leading to the production of ATP (Mitchell et al. (1976), Mitchell
et al. (1990)). The other fundamental property of coenzyme Q10 is
as an antioxidant, neutralizing free radicals (Beyer et al. (1990),
Vittalba et al. (1997)).
##STR00001##
[0009] Coenzyme Q10 is a benzoquinone derivative flanked by a long
isoprene side chain, most often composed of ten isoprenoid units
(whence the name coenzyme Q10). As this coenzyme is not soluble in
water, it is only encountered in lipid membranes like the internal
membrane of the mitochondrion, where it can freely diffuse among
the membrane phospholipids.
[0010] Coenzyme Q10 can exist in three oxidation states: a reduced
form (CoQH2 or UQH2), an oxidized state (CoQ10), and an
intermediate form, the radical ubisemiquinone)(Q.sup.0).
[0011] The biosynthesis of coenzyme Q10 starts with thyrosine for
the quinone core and with farnesyl pyrophosphate for the side
chain. The enzyme responsible for this last reaction, which is an
essential step in the biosynthesis of coenzyme Q10, is transphenyl
transferase (or polyphenyl transferase).
[0012] The search for compounds capable of stimulating the
synthesis of coenzyme Q10 or the energy synthesis of ATP, and/or
protecting the cells from damage caused by free radicals is an
important concern of medical research and of cosmetics. Solutions
have thus been proposed such as the intake of substances of peptide
origin exhibiting antioxidant properties (WO2005097060,
JP2006131626) or vitamin C (US 2004/0086526) or L-ergothionine (WO
9836748).
[0013] The present invention has as its principal objective the use
of a peptide active principle originating from the hydrolysis of
amaranth of the species Amaranthus hypochondriacus, capable of
protecting the skin from external aggressions and of combating
cutaneous aging. The said active principle could be used alone or
in association with at least one other active principle. The
inventors have highlighted a therapeutic activity, notably a
dermatological and cosmetic one, of such an active principle. It
has been particularly brought out that this active principle, when
applied on the skin, has a strong protective action against
oxidative damage to which the skin is subjected and promotes the
synthesis of ATP in a significant way, as well as the synthesis or
the activity of the enzyme transphenyl transferase and of coenzyme
Q10. This new active principle, capable of increasing cellular
energy and of protecting the skin from oxidative damage thus opens
up new therapeutic and cosmetic perspectives.
[0014] An "active principle capable of increasing cellular energy
and of protecting the skin from oxidative damage" is understood to
be any substance of vegetable origin, and more particularly
originating from amaranth of the species Amaranthus hypochondriacus
capable of increasing the synthesis of intracellular ATP and of
exhibiting protective properties in cells or tissues subjected to
an oxidative stress of physico-chemical or environmental origin.
The active principle capable of increasing cellular energy and of
protecting the skin from oxidative damage is, according to the
invention, an extract of peptide nature and originates from the
hydrolysis of proteins of amaranth of the species Amaranthus
hypochondriacus. "Of peptide nature" is understood to be a mixture
of compounds represented by peptides of polypeptides.
[0015] The term "peptide" designates a chaining of two or several
amino acids linked to one another by peptide bonds or by modified
peptide bonds, the term "polypeptide" designating a peptide of
greater size.
[0016] The expression "biologically active" is understood to be
"that posses an in vivo or in vitro activity characteristic of the
activity of the active principle according to the invention".
[0017] The term "hydrolysate or originating from the hydrolysis"
designates any substance or mixture of substances, or isolated
preparation, obtained after hydrolysis of vegetable matter.
[0018] The peptide active principle according to the invention is
obtained by the extraction of proteins of vegetable origin, after a
controlled hydrolysis that releases biologically active peptide
fragments.
[0019] A great many proteins found in plants are likely to contain
these sequences within their structure. Controlled hydrolysis
enables these peptide fragments to be released. It is possible, but
not necessary to achieve the invention, either to extract the
proteins concerned first and then to hydrolyze them, or to perform
the hydrolysis first on a raw extract and to subsequently purify
the peptide fragments. It is also possible to use certain
hydrolyzed extracts without purifying the peptide fragments in them
which correspond to peptides with the general formula (I) according
to the invention, but at the same time ensuring the presence of the
said fragments by appropriate analytical means.
[0020] In order to achieve the extraction, the entire plant may be
used or a specific part of the plant (leaf, berry, etc.).
[0021] The amaranth is an annual plant in the family Amaranthaceae
belonging to the genus Amaranthus, some species of which are
cultivated as ornamental plants for their flowers with a
spectacular ear, and sometimes as culinary plants, for their edible
leaves, like spinach greens or for their seeds.
[0022] According to the invention, the species used is Amaranthus
hypochondriacus, the vegetable material used is the seed, and
preferentially the seed separated from its husk by a dehusking
step.
[0023] In a first step, the plant is crushed using a plant crusher.
The powder thus obtained may ultimately be "delipidated" with the
aid of a traditional organic solvent (like, for example, alcohol,
hexane, or acetone).
[0024] Then the extraction of proteins from the plane is performed
according to the modified traditional procedure (Osborne (1924);
the plant crush is placed in suspension in an alkaline solution
containing an adsorbent product of the insoluble polyvinyl
polypyrrolidone (PPVP) type (0.01-20%). Indeed, it has been
observed that only operations of hydrolysis and final purification
are facilitated by this means. The concentration of substances of
the phenolic type which interact with the proteins are thus found
to be reduced.
[0025] The soluble fraction is collected after the centrifuging and
filtering steps, this raw solution then constituting as first form
of the extract, containing the proteins, the glucides, and possibly
lipids.
[0026] The proteins are then precipitated, varying the ionic
strength when acidifying the medium, which enables the soluble
components and the nucleic acids to be eliminated.
[0027] The precipitate is then washed with an organic solvent such
as, for example, ethanol or methanol, and the solvent is evaporated
by drying in a vacuum. The precipitate, rich is proteins, is placed
in solution again in water or another solvent and then consists of
a more purified form of the hydrolysate.
[0028] The extraction may also be performed in a neutral or acidic
medium, always in the presence of polyvinyl polypyrrolidone. After
a filtration step, the precipitation step is them performed using a
traditional precipitation agent such as the salts (sodium chloride,
ammonium sulfate) or an organic solvent (alcohol, acetone). The
precipitate obtained may be separated from the agents of
precipitation by dialysis after being placed again in solution in
water or another solvent.
[0029] The proteinic fraction isolated according to the invention
is then hydrolyzed under controlled conditions to generate soluble
polypeptides and peptides. Hydrolysis is defined as a chemical
reaction implying the cleaving of a molecule by water, this
reaction being able to be performed in a neutral acidic, or basic
medium. According to the invention, the hydrolysis is achieved by
chemical means and/or in an advantageous manner by proteolytic
enzymes. The use of endoproteases of vegetable origin may then be
cited (papayine, bromelaine, ficine) and of micro-organisms
(Aspergillus, Rhizopus, Bacillus, etc.).
[0030] For the same reasons as before, during this step of
controlled hydrolysis, a quantity of polyvinyl pyrrolidone is added
to the reaction medium. After filtering, the solution obtained
constitutes the active hydrolysate. The active hydrolysate may then
be purified in order to select the molecular weights and the nature
of the peptides generated. Fractionation may be performed to
advantage by ultrafiltration and/or by a chromatographic-type
method.
[0031] Any one of the more or less purified forms of the
hydrolysate is then solubilized in water or in any mixture
containing water, and then sterilized by ultrafiltration.
[0032] The vegetable hydrolysate obtained according to the
invention is analyzed qualitatively and quantitatively for its
physico-chemical characteristics and its content in compounds of a
protein and peptide nature. Compounds of a peptide nature are
understood to be the protein fragments, peptides, and free amino
acids present in the mixture. The peptides, amino acids, and
protein fragments are measured according to traditional techniques,
well know to the professional.
[0033] Thus, according to an advantageous embodiment of the
invention, the active vegetable hydrolysate has a pH between 4 and
7, and preferentially between 5 and 6, a dry extract titering
between 1 and 8 g/l, and preferably between 2 and 5 g/l. Its
content in compounds of a peptide nature is between 0.12 and 5 g/l,
and preferentially between 0.5 and 2 g/l, and its content in sugars
is from 0.5 to 2.5 g/1.
[0034] According to one advantageous embodiment of the invention,
the active principle according to the invention is solubilized in
advance in one or several solvents traditionally used by the
professional, such as water, glycerol, ethanol, propylene glycol,
butylene glycol, dipropylene glycol, ethoxylated or propoxylated
diglycols, cyclic polyols, Vaseline, a vegetable oil, or any
mixture of these solvents.
[0035] According to yet another advantageous embodiment of the
invention, the active principle according to the invention is
solubilized in advance in a cosmetic or pharmaceutical vehicle like
the liposomes or adsorbed onto powdered organic polymers or mineral
supports like the talcs and bentonites, and is more generally
solubilized in, or fixed upon, any cosmetically or pharmaceutically
acceptable vehicle.
[0036] The compositions according to the invention could be applied
in any appropriate way, particularly oral, parenteral, or
externally topical, and their formulation will be adapted by the
professional, in particular for cosmetic or dermatological
compositions. Advantageously, the compositions according to the
invention are intended for administration by topical, cutaneous
means. These compositions shall therefore contain a cosmetically
and/or dermatologically acceptable medium, that is, compatible with
the skin and the appendages, and they cover all the cosmetic or
dermatological forms. These compositions could, in particular, be
in the form of creams, oil-in-water or water-in-oil emulsions or
multiple emulsions, solutions, suspensions, gels, milks, lotions,
sticks, or even powders, adapted to an application onto the skin,
the lips, and/or the appendages.
[0037] These compositions include the excipients necessary for
their formulation, such as solvents, thickeners, diluents,
surfactants, antioxidants, colorants, preservatives, or
perfumes.
[0038] Of course, the professional will take care to choose
possible supplemental compounds, active or not, and/or their
quantity, such that the advantageous properties of the mixture are
not altered by the addition envisioned.
[0039] The composition usable according to the invention can, in
particular, consist of a composition for hair care, and
particularly a shampoo, a conditioner, a blow-dry lotion, a
treatment lotion, a cream or a styling gel, a restructuring lotion
for the hair, a mask, etc. The cosmetic composition according to
the invention can be used particularly in treatments implementing
an application that is followed or not by a rinse, or even in the
form of shampoo.
[0040] It can also come in the form of a dye or a mascara to be
applied with brush or comb, in particular on the eyelashes,
eyebrows, or hair.
[0041] Advantageously, the usable compositions contain, in
addition, at least one other active agent promoting the action of
the peptide active principle according to the invention. Thus, the
composition according to the invention may associate, with the
active principle, capable of increasing cellular energy and of
protecting the skin from oxidative damage, active agents having an
antioxidant action, or else stimulating the synthesis of dermal
macromolecules, or else stimulating energy metabolism. For example,
as an active principle having an anti-radical or antioxidant
action, vitamin C, vitamin E, or coenzyme Q10, or the polyphenolic
extracts of plants may be cited. Active anti-radical principles are
understood to be any compound capable of trapping free radicals.
These active principles are capable of blocking the chain reactions
of free radicals before the final stages of degradation of the
biological constituents of the skin, and because of this they have
an antioxidant activity.
[0042] As an active agent stimulating the synthesis of dermal
macromolecules (laminin, fibronectin, collagen), the collagen
peptide marketed under the name of Collaxyl.RTM. by the Vincience
Company may be cited.
[0043] Finally, the active principle marketed under the name of
GP4G.RTM. by the Vincience Company may be cited as an active agent
stimulating energy metabolism.
[0044] From another angle, the composition according to the
invention may be a sun-related composition, that is, a composition
that aids in protection against solar radiation. Thus, there may be
advantageously added to the composition according to the invention
active agents aiding in solar protection such as, for example,
solar filters.
[0045] It is quite obvious that the invention is directed toward
mammals in general and more particularly toward human beings.
[0046] The effective quantity of active principle corresponds to
the quantity of amaranth hydrolysate obtained according to the
invention necessary to obtain the result sought, that is to say, to
increase the synthesis of ATP, to protect the skin from oxidative
damage, and more generally to protect the skin from external
aggressions and to prevent or treat cutaneous aging.
[0047] According to an advantageous embodiment of the invention,
the active principle originating from amaranth is present in the
compositions of the invention in a concentration between
approximately 0.0001 and 20%, and preferentially in a concentration
between approximately 0.05 and 5%, relative to the total weight of
the final composition.
[0048] These compositions could come particularly in the form of an
aqueous, hydroalcoholic, or oily solution, an oil-in-water or
water-in-oil emulsion, or multiple emulsions. They can also come in
the form of creams, suspensions, or even powders, adapted to
application onto the skin, the mucous membranes, the lips, and/or
the appendages. These compositions can be more or less fluid and
have the appearance of a cream, a lotion, a milk, a butter, an
ointment, a gel, a paste, or a mousse. They can also come in solid
form like a stick or be applied on the skin in the form of an
aerosol. They can be used as a care product and/or as a makeup
product for the skin.
[0049] These compositions include, in addition, any additive
commonly used in the application domain envisioned, as well as the
adjuvants necessary for their formulation, such as solvents,
thickeners, diluents, antioxidants, colorants, solar filters,
self-bronzing agents, pigments, vehicles, preservatives, perfumes,
odor absorbents, active cosmetic or pharmaceutical agents,
essential oils, vitamins, essential fatty acids, surfactants,
film-forming polymers, etc.
[0050] In any case, the professional will take care that these
adjuvants, as well as their proportions, are chosen in such a way
as not to harm the advantageous properties sought in the
composition according to the invention. These adjuvants can, for
instance, correspond to 0.01 to 20% of the total weight of the
composition. When the composition of the invention is an emulsion,
the fatty phase may represent 5 to 80% by weight and preferably 5
to 50% by weight relative to the total weight of the composition.
The emulsifiers and co-emulsifiers used in the composition will be
chosen from among those traditionally used in the domain
considered. For example, they may be used in a proportion of 0.3 to
30% by weight relative to the total weight of the composition.
[0051] By means of its particular activities, the active principle
according to the invention could be used advantageously in a
cosmetic composition or for the preparation of a pharmaceutical
composition.
[0052] In particular, the active principle according to the
invention could be used advantageously in a cosmetic composition
intended to combat in a preventive and/or curative manner the
manifestations of cutaneous aging and, more specifically, to combat
and/or prevent photo-induced aging (photo-aging). Cutaneous
manifestations of aging are understood to be any alteration of the
external appearance of the skin due to aging, such as, for example,
wrinkles and fine wrinkles, shriveled skin, flabby skin, thin skin,
lack of elasticity and/or tonus of the skin, dull skin without a
glow or pigmentation spots on the skin, as well as any internal
alteration of the skin that is not manifested systematically in an
altered external appearance, such as, for example, any internal
degradation of the skin following an exposure to ultraviolet (UV)
rays. The active principle according to the invention, or the
composition containing it, will enable combating, in particular,
the loss of elasticity and firmness of the skin.
[0053] The use of the peptide, or of a composition containing it,
will allow the skin and the appendages to be protected and to
better resist environmental stresses. Thus, an essential aspect of
the invention is the use of the active principle according to the
invention, in a cosmetic composition for protecting the skin and
the appendages against oxidative damage, thanks to a protective
activity with respect to reactive species of oxygen, the said
active principle being used to advantage as an active antioxidant
principle, and/or as an active anti-radical principle, and/or as an
active antiglycation principle. An active anti-radical principle is
understood to be any compound capable of trapping free radicals
before the final stages of degradation of the biological
constituents of the skin, and they are then called antioxidant
compounds. An active antiglycation principle is understood to be
any compound capable of limiting the cellular damage caused by
glycation or glycoxidation reactions. Thus, the active principle
according to the invention will enable combating the aesthetic
damage caused to the skin and/or the hair by free radicals.
[0054] Also, the active principle may be used advantageously in a
cosmetic composition to protect the skin and the appendages against
all types of external aggressions. The expression "external
aggressions" is understood to be aggressions which the environment
can produce. By way of example, aggressions may be cited such as
pollution, UV rays, or even products of an irritating nature such
as surfactants, preservatives, and perfumes. Pollution is also
understood to be both "outdoor" pollution due, for example, to
Diesel-fuel particles, ozone, or heavy metals, as well as "indoor"
pollution, which may be due, in particular, to the emissions of the
solvents of paint, glue, or wallpaper (such as toluene, styrene,
xylene, or benzaldehyde), or even cigarette smoke.
[0055] The active principle according to the invention may be
advantageously used in a cosmetic composition or for preparing a
pharmaceutical composition, as an active photo-protective principle
and, more particularly, as a so-called "secondary" active
photo-protective principle. Primary active photo-protective
principles are, in effect, differentiated from secondary active
photo-protective principles. Primary active photo-protective
principles are substances that exercise a physical power: they are
capable of absorbing UV rays and releasing them in the form of heat
in order to protect the skin. Secondary active photo-protective
principles are substances that generally have a biological effect;
they are, for example, active principles capable of limiting the
damage caused to the DNA and to the membranes by UV rays
penetrating into the skin.
[0056] The invention also has, as an object, use in a cosmetic
composition of an effective quantity of active principle as
previously described to prevent damage caused to the skin by
exposure to the sun or exposure to ionizing radiation during
radiotherapy.
[0057] The invention also has, as an object, use in a cosmetic
composition of an effective quantity of active principle as
previously described to increase the synthesis of intracellular ATP
in the cells of the skin.
[0058] The invention also has, as an object, use in a cosmetic
composition of an effective quantity of active principle as
previously described to increase the activity or the synthesis of
the enzyme transphenyl transferase and/or coenzyme Q10 in the cells
of the skin.
[0059] The invention again is related to use in a cosmetic
composition of an effective quantity of active principle as
described previously to protect the skin from damage caused by free
radicals.
[0060] The invention consists too of the use of an effective
quantity of active principle, as described previously, for making a
pharmaceutical composition intended to mitigate a pathology linked
to oxidation processes or even certain pathologies of aging.
[0061] Finally, the invention again consists of a
cosmetic-treatment procedure intended to protect the skin and the
appendages from external aggressions and to combat cutaneous aging,
characterized by the application onto the skin or the appendages to
be treated of a composition containing an effective quantity of
active principle according to the invention.
[0062] Specific embodiments of this cosmetic-treatment procedure
also result from the preceding description. Other advantages and
characteristics of the invention will be more apparent upon reading
the examples given as illustration and non-restrictive.
EXAMPLE 1
Preparation of Active Principle Starting with Amaranth (Amaranthus
hypochondriacus)
[0063] The active agent is obtained starting from an extract of
plants of the species Amaranthus hypochondriacus.
[0064] In a first step, 1 kg of hulled amaranth seeds is crushed in
a cereal crusher and the meal obtained is delipidated by means of
the action of an organic solvent, hexane. After filtration and
drying in a vacuum, the powder obtained is placed in suspension in
an aqueous alkaline solution (dilution to 1/10) of pH 10,
containing 1% polyvinyl polypyrrolidone (Polyclar V ISP). This
mixture is kept while stirring for a time sufficient to allow
solubilization of the soluble fractions. The extraction temperature
is variable (between 4 and 80.degree. C.). Preferentially, the
operation is carried out cold. After this extraction phase, the
medium is clarified by centrifuging and then filtered on a plate
filter. This filtrate, which contains the soluble fractions of the
amaranth, is then subjected to protein precipitation under varying
ionic strength in a neutral or acidic medium, which enables the
soluble glucide components, the lipids, and the nucleic acids to be
eliminated. The medium is brought to a pH of 3.5. The supernatant
is eliminated and the precipitate is then washed using a solvent
such as, for example, ethanol or methanol, and then the solvent is
evaporated by drying in a vacuum.
[0065] At this stage, about 50 grams of clear, yellow-colored
powder of raw proteinic extract are obtained, containing:
[0066] Proteins: 75%
[0067] Glucides: 20%
[0068] Lipids: 5%
The precipitate, rich in proteins is placed back into solution in
water or another solvent.
[0069] The raw proteinic extract is then subjected to a series of
controlled and selective hydrolyses consisting of chemical and
enzymatic hydrolyses in the presence of 5% PVPP (Polyclar V) and
endopeptidases to cysteine (papayine, ficine). After reaction, the
hydrolysate is filtered onto a plate and then onto a sterilizing
cartridge (0.2 .mu.m).
[0070] A clear-colored hydrolysate is then obtained titering from
15 to 30 g/l of dry extract, which is then diluted such that the
composition in compounds of a peptide nature determined by Lowry's
method is between 0.1 and 5 g/l, and preferentially between 0.5 and
2 g/l. The physico-chemical analysis of the vegetable hydrolysate,
which constitutes the active principle, shows that its pH is
between 4 and 7, and preferentially between 5 and 6, the dry
extract titers between 1 and 8 g/l, and preferably between 2 and 5
g/l. Its content in compounds of a peptide nature is between 0.1
and 5 g/l, and preferentially between 0.5 and 2 g/l, and its
content in sugars is from 0.5 to 2.5 g/1.
EXAMPLE 2
Preparation of Active Principle Starting with Amaranth (Amaranthus
hvpochondriacus)
[0071] A variant of the protocol of example 1 consists of carrying
out the same sequence of controlled and selective enzymatic
hydrolyses, but in the presence of 0.5% PVPP. A clear-colored
hydrolysate is then obtained titering 15 to 30 g/l of dry extract
after a sterilizing filtration.
[0072] You then proceed to ultrafiltration of the solution onto a
Helicon millipore filtration cartridge (1 kD cutoff). The high
molecular weights contained in the retentate are isolated, and the
filtrate is retained.
[0073] The concentration in compounds of a peptide nature
determined by Lowry's method to be between 0.1 and 5 g/l, and
preferentially between 0.5 and 2 g/l. The physico-chemical analysis
of the vegetable hydrolysate, which constitutes the active
principle, shows that its pH is between 4 and 7, and preferentially
between 5 and 6, the dry extract titers between 1 and 8 g/l, and
preferably between 2 and 5 g/l. Its content in compounds of a
peptide nature is between 0.1 and 5 g/l, and preferentially between
0.5 and 2 g/l, and its content in sugars is from 0.5 to 2.5
g/1.
[0074] Another variant consists of carrying out a purification of
the active principle obtained according to example 1 or 2, by
ion-exchange chromatography, on a gel TSK column (TsosHaas) with a
pH 7 phosphate buffer.
EXAMPLE 3
Disclosure of the activator effect of the active principle
according to example 1 on the synthesis of intracellular ATP
[0075] The aim of this study was to determine the effect of the
active principle on the synthesis of ATP.
[0076] Protocol
[0077] This study was conducted using an ATP Bioluminescence Assay
Kit HS II (Roche Applied Science). Dermal fibroblasts were treated
with a 1% solution of the active principle according to example 1
for a period of 1 to 3 hours. At the end of the incubation time,
the tubes were emptied of their medium and rinsed with 2 ml of cold
PBS before adding 250 .mu.l of a lysing buffer provided in the kit.
The cells of each tube were then scraped out and collected in 14-ml
tubes. Each tube was rinsed with 2.times.500 .mu.l of cold PBS and
the whole was collected again in the respective tubes. Starting
with these samples, a dilution to 1/12000th was achieved in cold
PBS before each reading. ATP measurement was performed on these
samples: 50 .mu.l of this dilution was placed in a Luma cuvette and
50 .mu.l of luminol was added. After 10 seconds, the luminescence
reading was begun. The values were standardized relative to the
quantity of proteins for each sample. The measurements were made
using the Biocounter M2010A Luma.RTM./3M equipment.
[0078] Results
[0079] The ATP measurements show that there is an increase of 17%
in the amount of intracellular ATP after one hour and 67% after 3
hours of culture in the cells treated with the active principle,
compared to the untreated cells.
[0080] Conclusion
[0081] The active principle strongly activates the energy level of
synthesis of intracellular ATP in cutaneous cells.
EXAMPLE 4
Evaluation of the Protective Effect of the Active Principle
According to Example 1 with Respect to Oxidative Damage
[0082] The aim of this study was to determine the protective effect
of the active principle according to example 1 with respect to
dermal fibroblasts subjected to an oxidative stress caused by UVB
rays or hydrogen peroxide (H.sub.2O.sub.2). In order to evaluate
the oxidative damage undergone by the cells, measurements were made
of protein carbonylation.
[0083] The carbonylation of proteins results from the oxidative
cleaving of proteins or from an oxidation of the residues of
arginine, lysine, proline, or threonine. Protein carbonylation
measurement was done using the (enzyme immunoassay (EIA) technique
Protocol
[0084] Fibroblasts in culture had been placed in the presence of 1%
active principle 72 hours before, during, and after the oxidative
stress (UVB irradiation at 50 mJ/cm.sup.2 or treatment with 2 mM of
H.sub.2O.sub.2). Controls untreated and not subjected to the
oxidative stress were achieved.
[0085] The carbonylation measurement consisted of using DNP
(dinitrophenyl), which has the property of being specifically fixed
on the carbonyl groups of proteins. Fixed DNP is then measured by
an ELISA method, thank to an anti-DNP antibody coupled to a
peroxydase. An oxided bovine serum albumin (BSA) scale (which
recognizes the concentration in carbonyl groups) is used for
standardization.
[0086] Results
[0087] The results obtained show a reduction of 30% in protein
carbonylation when the cells were treated with the active principle
according to example 1, compared with untreated cells.
[0088] More particularly, a reduction of 20% is observed in the
carbonylation when the cells treated with the active principle
according to example 1 are subjected to UVB irradiation or to an
oxidative stress with H.sub.2O.sub.2, compared with cells
irradiated or stressed but not treated with the active
principle.
[0089] Conclusions
[0090] The active principle according to example 1 effectively
protects the cutaneous cells against oxidative damage caused by UVB
rays or hydrogen peroxide.
EXAMPLE 5
Evaluation of the Protective Effect of the Active Principle
According to Example 1 with Respect to a Stress Induced by
Glycation
[0091] The aim of this study was to determine the protective effect
of the active principle according to example 1 with respect to an
ex vivo epidermal culture subjected to a stress by a glycant
agent.
[0092] Protocol
[0093] Biopsies of human skin were kept in an ex vivo culture,
treated with a 1% solution 24 hours before and then 24 hours after
being placed in the presence of a glycant agent (5 or 10 mM methyl
glyoxal). Slices and histological hematoxyline-eosine (H&E)
stainings enabled the quality of the cutaneous structures to be
evaluated.
[0094] Results
[0095] Observation shows a clear reduction of the signs of cellular
stress and better preservation of cutaneous structures in the skin
biopsies treated with the active principle according to example 1,
compared with untreated skin biopsies.
[0096] Conclusions
[0097] The active principle according to example 1 protects the
skin from a stress induced by glycation.
EXAMPLE 6
Preparation of Compositions
[0098] 1. Sunscreen cream
TABLE-US-00001 International Nomenclature of Cosmetic Ingredients
Trade name (INCI) names % W/W PHASE A Demineralized water Aqua
(water) In sufficient quantity Pemulen .RTM. TR-1 Acrylates/C10-30
alkyl 0.40 acrylate cross-polymer Glycerine Glycerin 3.00 Nipastat
.RTM. Sodium Sodium methylparaben (and) 0.15 sodium ethylparaben
(and) sodium butylparaben (and) sodium propylparaben (and) sodium
isobutylparaben PHASE B Parsol .RTM. MCX Ethylhexyl
methoxycinnamate 7.50 Eusolex .RTM. 4360 Benzophenone-3 3.00 Parsol
.RTM. 1789 Butyl methoxydibenzoyl- 2.00 methane Myritol .RTM. 318
Caprylic/capric triglyceride 4.00 Emulgade .RTM. SEV Hydrogenated
palm glycerides 5.00 (and) ceteareth-20 (and) ceteareth-12 (and)
cetearyl alcohol Propylparaben Propylparaben 0.15 Nacol .RTM. 16-98
Cetyl alcohol 1.00 PHASE C TEA Triethanolamine 0.20 PHASE D Active
principle 3 acc. to ex. 1 Perfume Perfume (fragrance) In sufficient
quantity Colorant In sufficient quantity
The constituents of phase A and of phase B are heated separately to
a temperature between 70.degree. C. and 75.degree. C. Phase B is
emulsified in phase A while stirring. Phase C is added, at
45.degree. C., while increasing the stirring. Phase D is then added
when the temperature was below 40.degree. C. Cooling is continued
down to 25.degree. C. with brisk stirring.
[0099] 2. After-Sun Milk
TABLE-US-00002 Trade name INCI names % W/W PHASE A Montanov .TM. L
C14-22 alcohols (and) C12- 3.00 20 alkyl glucoside Waglinol 2559
Cetearyl isononanoate 4.00 Tegosoft .RTM. TN C12-15 alkyl benzoate
3.00 Apricot kernel oil Prunus armeniaca (apricot) 2.00 kernel oil
Avocado oil Persea gratissima (avocado) 1.00 oil Abil .RTM. 350
Dimethicone 1.00 PHASE B Demineralized water Aqua (water) In
sufficient quantity PHASE C Simulgel .TM. EG Sodium
acrylate/acryloyl- 0.4 dimethyl taurate copolymer (and)
isohexadecane (and) polysorbate 80 copolymer (and) polysorbate 80
PHASE D Phenonip .RTM. Phenoxyethanol (and) 0.30 methylparaben
(and) ethylparaben (and) butylparaben (and) propylparaben (and)
isobutylparaben ethylparaben and propylparaben and butylparaben
Germall .RTM. 115 Imidazolidinyl urea 0.20 PHASE E Active principle
0.1 acc. to ex. 1
Prepare phase A while stirring. Incorporate the xanthan gum
gradually, with dispersant stirring. Phases C and D will be
incorporated once the gel has set. Phase E, prepared in advance to
the point of perfect DHA dissolution, will then be added. Adjust
the pH, if necessary, to 4-4.5. Color and perfume.
[0100] 3. Anti-Aging Cream
TABLE-US-00003 Trade name INCI names % W/W PHASE A Montanov .TM. 68
Cetearyl alcohol (and) 6.00 cetearyl glucoside Squalane Squalane
3.00 Cetiol .RTM. SB 45 Butyrospermum parkii (Shea 2.00 butter)
Waglinol 250 Cetearyl ethylhexanoate 3.00 Amerchol L-101 Mineral
oil (and) lanolin 2.00 alcohol Abil .RTM. 350 Dimethicone 1.50 BHT
Butylhydroxytoluene 0.01 Coenzyme Q10 Ubiquinone 0.10 PHASE B
Avocado oil Persea gratissima (avocado) 1.25 oil Phenonip .RTM.
Phenoxyethanol (and) 0.75 methylparaben (and)\ ethylparaben (and)
butylparaben (and) propylparaben (and) isobutylparaben PHASE C
Demineralized water Aqua (water) In sufficient quantity Butylene
glycol Butylene glycol 2.00 Glucam .TM. E10 Methyl gluceth-10 1.00
Allantoin Allantoin 0.15 Carbopol .RTM. Ultrez 10 Carbomer 0.20
PHASE D TEA Triethanolamine 0.18 PHASE E Active principle 0.5 acc.
to ex. 1 GP4G .RTM. Water (and) Artemia extract 1.50 Collaxyl .RTM.
Water (and) butylene glycol 3.00 (and) hexapeptide-9 PHASE F
Perfume Perfume (fragrance) In sufficient quantity Colorant In
sufficient quantity
Prepare and melt phase A at 65-75.degree. C. Heat phase C to
65-70.degree. C. Phase B is added to phase A just before
emulsifying A in B. At about 45.degree. C., the carbomer is
neutralized by the addition of phase D. Phase E is then added with
light stiffing, and cooling is continued to 25.degree. C. Phase F
is then added, if desired.
[0101] 4. Daytime Protection Cream
TABLE-US-00004 Trade name INCI names % W/W PHASE A Emulium delta
.RTM. Cetyl alcohol (and) glyceryl 4.00 stearate (and) PEG-75
stearate (and) ceteth-20 (and) steareth-20 Lanette O Cetearyl
alcohol 1.50 DC 200 Fluid .RTM./100 cs Dimethicone 1.00 DUB 810C
Coco caprylate/caprate 1.00 DPPG Propylene glycol 3.00
dipelargonate DUB DPHCC Dipentaerythrityl 1.50
hexacaprylate/hexacaprate Cegesoft .RTM. PS 6 Vegetable oil 1.00
Vitamin E Tocopherol 0.30 Phenonip Phenoxyethanol (and) 0.70
methylparaben (and) ethylparaben (and) butylparaben (and)
propylparaben (and) isobutylparaben PHASE B Demineralized water
Aqua In sufficient quantity, 100 Glycerine Glycerin 2.00 Carbopol
.RTM. ETD 2020 Acrylates/C10-30 alkyl 0.15 acrylate cross-polymer
Keltrol CG-BT Xanthan gum 0.30 PHASE C Sodium hydroxide (10% Sodium
hydroxide 0.30 solution) PHASE D Demineralized water Aqua 5.00
Stay-C .RTM. 50 Sodium ascorbyl phosphate 0.50 PHASE E Butylene
glycol Butylene glycol 2.00 Dekaben CP Chlorphenesin 0.20 PHASE F
GP4G .RTM. Water (and) Artemia extract 1.00 Active principle 5 acc.
to ex. 1
Prepare phase A and heat to 75.degree. C. while stiffing. Prepare
phase B while dispersing the Carbopol.RTM. and then the xanthan gum
while stirring. Let rest. Heat to 75.degree. C. At temperature,
emulsify A in B with rotor-stator stirring. Neutralize with phase C
while stirring rapidly. After cooling to 40.degree. C., add phase D
and then phase E. Cooling is continued with light stirring and
phase F is added.
* * * * *